Epitope-tagged insulin-like growth factor-I expression in muscle

Development of a recombinant insulin like growth factor I (IGF-I) that is distinguishable from its endogenous counterpart would provide a powerful tool for delineating the role of IGF in myogenesis. Therefore, the objective of this study was to create an epitope-tagged IGF-I that retains biological activity and determine whether expression of this construct is possible in muscle tissue following direct DNA injection. Expression vectors were created that encoded porcine IGF-I containing a T7 (11-amino acid) epitope-tag (TIGF). Immunoreactivity of the purified recombinant TIGF was confirmed using monoclonal antibodies. Biological activity was evaluated by examining differentiation of myoblasts cultured with TIGF or transfected with TIGF plasmid DNA. Addition of purified TIGF to myoblast cultures stimulated (P < 0.05) muscle creatine kinase levels similar to insulin (10(-5) M). Likewise, transfection of L6A1 with TIGF DNA hastened (P < 0.01) differentiation compared to control pcDNA-transfected myoblasts. The integrity of the recombinant protein was confirmed using a sandwich-configured enzyme linked immunosorbent assay. Finally, recombinant TIGF DNA was injected in porcine muscle and the ability to detect TIGF protein was evaluated. TIGF expression was detected in muscle fibers of injected porcine muscle. These data show that a T7 amino acid tag placed on the amino terminus of the IGF-I protein remains intact during processing and does not interfere with the biological activity of the molecule. Use of this DNA construct is an excellent tool for investigating the role of IGFs in control muscle development and provides a model to investigate other regulators of animal growth.     

Evaluation of intratesticular injection of glycerol for nonsurgical sterilization of dogs

OBJECTIVE: To determine efficacy of intratesticular injection of glycerol as a means to sterilize dogs. ANIMALS: Six 2- to 4-year-old sexually intact male dogs. PROCEDURE: Testicular volume, serum testosterone concentration, and number of sperm in ejaculate were determined before and at weekly intervals for 12 weeks following intratesticular injection of a 70% glycerol solution (1 ml/testis). Testicular tissue specimens for histologic evaluation were obtained 4, 8, and 12 weeks following injection. RESULTS: Number of sperm in ejaculate following glycerol injection was not significantly different from number before injection. Compared with preinjection values, serum testosterone concentrations were significantly less 5, 9, and 10 weeks following injection, although libido and ejaculate characteristics were unchanged. Testicular swelling was evident 24 to 48 hours following injection, and testicular volume decreased starting on week 5 following treatment. Seminiferous tubules and evidence of normal spermatogenesis were detected in testicular biopsy specimens obtained from all dogs 12 weeks following injection. CONCLUSION AND CLINICAL RELEVANCE: Intratesticular injection of a 70% glycerol solution did not result in azoospermia and sterility in dogs.     

Stress and feather pecking in laying hens in relation to housing conditions

1. Possible association between high rates of feather pecking and increased stress were investigated in laying hens. 2. From week 19 to week 30 after hatching, 16 groups of 11 hens (white Lohman Selected Leghorn hybrids) were kept in pens with or without long-cut straw as foraging material and provided with food in the form of pellets or mash. 3. Stress was assessed by egg production, weight gain, tonic immobility (TI), heterophil/lymphocyte (H/L) ratio and antibody titres to sheep red blood cells (SRBC), tetanus toxoid (TT) and human serum albumin (HSA). 4. Provision of foraging material and food form influenced feather pecking. Rates of feather pecking were highest in groups housed without straw and fed on pellets. 5. Egg production was reduced in pens without straw but not affected by food form. Both the duration of TI and H/L ratios were influenced by provision of foraging material and food form. TI was longer and H/L ratios were increased in hens housed without straw and in those fed on pellets. Antibody titers to SRBC and TT were lower in pens without straw than with straw but not influenced by food form. 6. In conclusion, foraging material and food form affected both feather pecking and indicators of stress, suggesting that feather pecking in laying hens is associated with stress.     

Morphology and amplitude values of the electrocardiogram of Spanish-bred horses of different ages in the Dubois leads system

The aim of this work was to record the modifications of the form and amplitude of the electrocardiographic tracings during growth using the Dubois system and to study its sensibility to these physiological changes. This work was carried out on 179 healthy, young and adult Spanish-bred horses (98 females and 81 males). One electrocardiogram (ECG) was obtained from each horse using the Dubois leads system. The bifid shape deflection of the P wave was generally more frequent than the simple one in animals at 4 months of age or older. Amplitudes of the P2 component and of the simple positive wave presented significant differences (p < 0.01) between the age groups studied: animals up to 3 months in age and older than 2 years of age (highest amplitudes), and the rest of the animals (lowest amplitudes). Significant variations (p < 0.05) were found for the QS amplitude between animals of 1 month of age, and all the other groups. The multiple range analysis did not define a clear distribution of age groups for the amplitude of the T wave. It may therefore be concluded, that in the Spanish-bred horse up to 1 month of age, the form and amplitude of the QRS complex in the Dubois leads system differ from those of older animals. Thus, this study shows the sensibility of the Dubois leads system for detecting the electrocardiographic changes related to the growing process in the Spanish-bred horse. In addition, it provides standard values of electrocardiographic parameters in the Spanish-bred horse at different stages of growth.     

Pheasant ataxia: a new condition in pheasant poults

Between 1995 and 1997 a neurological condition in pheasant poults from 24 sites in England and Scotland was investigated. Affected birds showed varying degrees of ataxia and incoordinated movements and, in severe cases, recumbency, but generally remained alert with their heads held upright. The condition characteristically affected poults from seven weeks of age and the incidence on any one site was low. No significant bacteria were isolated consistently from brain tissue. The condition was characterised histologically by a non-suppurative meningoencephalitis, in which lesions were found predominantly in the cerebellum in 61 of 81 samples examined (75.3 per cent). A non-suppurative myelitis was recorded in 16 of 20 spinal cords examined. No lesions were recorded in peripheral neural tissue and lesions were rare in other tissues. The condition appeared not to have been recorded previously in pheasants. A viral aetiology was suspected but Newcastle disease virus was not involved.     

On the morphology of the liver in the two-humped camel (Camelus bactrianus)

The liver of a two-humped camel (Camelus bactrianus) was examined by means of gross anatomy and histology. The liver appeared characteristically as an enlarged triangle in visceral aspect, and showed the following lobation: the left lateral and medial lobes, the right lobe, the caudate lobe and the quadrate lobe. These findings were similar to those for the one-humped camel. Histologically, a distinct lobulation, a typical arrangement of the interlobular connective tissue, and the trias was confirmed.     

Effects of intravenous administration of formaldehyde on platelet and coagulation variables in healthy horses

OBJECTIVES: To assess safety and determine effects of IV administration of formaldehyde on hemostatic variables in healthy horses. ANIMALS: 7 healthy adult horses. PROCEDURE: Clinical signs and results of CBC, serum biochemical analyses, and coagulation testing including template bleeding time (TBT) and activated clotting time (ACT) were compared in horses given a dose of 0.37% formaldehyde or lactated Ringer's solution (LRS), IV, in a 2-way crossover design. In a subsequent experiment, horses received an infusion of 0.74% formaldehyde or LRS. In another experiment, horses were treated with aspirin to impair platelet responses prior to infusion of formaldehyde or LRS. RESULTS: Significant differences were not detected in any variable measured between horses when given formaldehyde or any other treatment. Infusion of higher doses of formaldehyde resulted in adverse effects including muscle fasciculations, tachycardia, tachypnea, serous ocular and nasal discharge, agitation, and restlessness. CONCLUSIONS AND CLINICAL RELEVANCE: Intravenous infusion of formaldehyde at doses that do not induce adverse reactions did not have a detectable effect on measured hemostatic variables in healthy horses.     

Radial carpal bone fractures in 15 dogs

Between 1977 and 1992, 15 adult dogs were presented to the Universities of Bristol, Glasgow, Liverpool and London with fractures of the radial carpal bone which had occurred without a known episode of violent trauma. Eleven of the dogs were boxers and the other breeds affected were a Pharaoh hound, rottweiler, springer spaniel and a greyhound. The condition was bilateral in three cases. Eleven dogs were treated surgically by cast immobilisation (four cases), fragment removal (one case), lag screw fixation (three cases), pan-carpal arthrodesis (one case) and lag screw reduction followed by pan-carpal arthrodesis (two cases). The remaining four dogs were treated conservatively with rest and tactical use of non-steroidal anti-inflammatory drugs. Follow-up periods ranged from two months to 11 years. Varying degrees of lameness, which tended to be exacerbated by exercise, persisted in all cases. The radial carpal bone appears to have at least three separate centres of ossification which eventually fuse; their planes of fusion correspond approximately to the two main fracture lines. These areas of fusion could be weak points within the radial carpal bone.     

Prevalence of enteric zoonotic organisms in cats

OBJECTIVE: To determine prevalence of enteric zoonotic organisms in cats in north-central Colorado. DESIGN: Prospective study. SAMPLE POPULATION: Serum and fecal samples from 87 cats with diarrhea, 106 cats without diarrhea, and 12 cats for which fecal consistency was unknown. PROCEDURES: Samples were obtained from client-owned cats and cats at a humane society shelter. Serum was assayed for feline leukemia virus antigen and antibodies against feline immunodeficiency virus, IgM antibodies against Toxoplasma gondii, and IgG antibodies against T gondii and Cryptosporidium parvum. Microscopic examination of unstained feces was performed after centrifugation in a zinc sulfate solution, thin fecal smears were stained with acid fast stain and examined for C parvum, and bacteriologic culture of feces was used to detect aerobic and anaerobic bacteria. RESULTS: Enteric zoonotic organisms were detected in feces from 27 of 206 (13.1%) cats and included C parvum (5.4%), Giardia spp (2.4%). Toxocara cati (3.9%), Salmonella enterica serotype Typhimurium (1.0%), and Campylobacter jejuni (1.0%); each organism was detected in samples from cats with and without diarrhea. Although differences between groups were not significant, a higher proportion of shelter cats (18.2%) had enteric zoonotic organisms than client-owned cats (10.1%). CONCLUSIONS AND CLINICAL RELEVANCE: Enteric zoonotic organisms were detected in feces of 13.1% of cats, suggesting that cats, particularly those in homes of immunocompromised humans, should be evaluated for enteric zoonotic organisms.